Porphyrin compounds

ABSTRACT

The present invention provides the porphyrin compound which is used for photodynamic diagnosis and/or treatment for animals, and the photodynamic diagnostic and/or therapeutic agent for animals.

TECHNICAL FIELD

[0001] The present invention relates to a porphyrin compound or apharmaceutically acceptable salt thereof used for photodynamic diagnosisand/or treatment of animals. The present invention also relates to aphotodynamic diagnostic and/or therapeutic agent comprising theporphyrin compound or a pharmaceutically acceptable salt thereof, whichis used for photodynamic diagnosis and/or treatment, especially, oftumor in animals.

BACKGROUND ART

[0002] As a new method of treatment for cancer, photodynamic diagnosisand therapy (PDT: Photodynamic Therapy) has stepped into the limelight.It is a method in which a certain type of porphyrin derivatives isadministered to a subject by, for example, intravenous injection toretain the porphyrin derivative in the target cancerous tissues in thesubject, followed by laser irradiation to cause selective destruction ofthe cancerous tissues. The therapy utilizes the two properties of aporphyrin derivative, i.e., selectivity for cancerous tissues andphotosensitivity.

[0003] The only porphyrin derivative currently used in PDT is porphymersodium. Porphymer sodium is a mixture compound of 2- to 6-polymercomprising an ether and/or ester of hematoporphyrin derivative.Porphymer sodium is known to cause temporary photosensitivity as anundesirable side effect when administered to human body, and further,selective distribution to cancerous tissues is not sufficient forpractical use, and therefore the problem of accumulation in normaltissues is confirmed.

[0004] Under the circumstances, a patient treated with porphymer sodiumis required to stay in the dark for a long period of time untilporphymer sodium is completely excreted from the body so that normalcells are not damaged by the photosensitizing action of porphymer sodiumaccumulated in normal tissues. However, since porphymer sodium shows aslow excretion rate from normal tissues, it sometimes causesphotosensitivity to last for more than six weeks.

[0005] In addition, PDT using porphymer sodium has a problem withtransmission of the light irradiated by laser through tissues. That is,porphymer sodium has a longest wavelength absorption end at 630 nm and amolar absorption coefficient is as small as 3,000. Since there are manycomponents present in a living body which prevent the transmission oflight, such as oxyhemoglobin and water, the light with wavelength of 630nm exhibits a poor transmission through tissues, which cannotsufficiently reach to deep sites, thus, PDT using porphymer sodium isonly intended for cancers developing in the surface layers of 5 to 10 mmdepth. The wavelength which is least damaging by the light absorption tothe components in a living body is in a range of 650 to 750 nm,therefore, photosensitizers for PDT having the longest wavelengthabsorption end within such range are most desirable.

[0006] Laser devices themselves also have problems. Dye lasers, whichare most commonly used at present, have a poor stability in performanceand therefore are difficult to handle in practical use. On the otherhand, titanium-sapphire lasers enable to facilitate the practice of PDTconsiderably. However, these types of lasers are limited in theexcitable wavelength to not less than 670 nm and not more than 600 nm,and therefore are not applicable to porphymer sodium which has anabsorption wavelength of near 630 nm.

[0007] Recently, semiconductor lasers (670 nm), which are applicable tocompounds exhibiting an absorption near 670 nm, have been developed, andquite recently OPO-YAG laser has been developed, which made it possibleto cover almost all visible wavelengths.

[0008] As mentioned above, photosensitizers currently used for PDT havevarious defects and therefore development of new agents without suchdefects is strongly desired. In an attempt to overcome those problems, aprophyrin compound which is a single compound and exhibits itsadsorption in a longer wavelength region (650-800 nm) has been proposedas a second generation agent for PDT.

[0009] Examples of such second generation agent includes amino-levulinicacid (ALA) which is a protoporphyrin precursor; asparthyl-chlorin e6 (Npe6) which is a chlorin derivative; benzoporphyrin derivative (BPD) andmethatetrahydroxyphenylchlorin (m-THPC), both of which are new type ofchlorin derivatives obtained by the structural conversion fromhemoglobin-derived porphyrins.

[0010] In addition, the present inventors proposed chlorin derivativesand the analogues thereof, e.g., an alkoxyiminochlonyl aspartic acidderivative (Japanese Patent Application Laid-open Nos. 5-97857 and9-124652), confirming that these compounds are useful asphotosensitizers for PDT.

[0011] On the contrary, in mammals' case, suffering from cancer has beena great problem not only in human beings but in animals, especially, inpet animals which are breeding in a house. For treating the cancer ofthese pet animals, same treatments for human being such as administeringanticancer agent or radiotherapy have been performed. Under thesecircumstances, the present inventors have studied to develop theeffective therapeutic methods for treatment of cancer of pet animals andconfirmed that among the alkoxyiminochlonyl aspartic acid derivatives,ethoxyiminochlonyl aspartic acid derivatives are useful asphotosensitizers for PDT in animals.

[0012] Therefore, it is an object of the present invention to provide aporphyrin compound or a pharmaceutically acceptable salt thereof usedfor photodynamic diagnosis and/or treatment of animals.

[0013] Furthermore, it is other object of the present invention toprovide a photodynamic diagnostic agent and/or therapeutic agentcomprising the porphyrin compound or a pharmaceutically acceptable saltthereof, especially for diagnosis and/or treatment of tumor in animals.

DISCLOSURE OF THE INVENTION

[0014] To solve the above-mentioned objects, one aspect of the presentinvention provides a porphyrin compound represented by the followingformula (I):

[0015] wherein Asp represents a residue of aspartic acid, or apharmaceutically acceptable salt thereof, used for photodynamicdiagnosis and/or treatment of animals.

[0016] Another aspect of the present invention provides a porphyrincompound represented by the following formula (II):

[0017] wherein Asp represents a residue of aspartic acid, or apharmaceutically acceptable salt thereof, used for photodynamicdiagnosis and/or treatment of animals.

[0018] Still another aspect of the present invention provides aphotosensitizer for the photodynamic diagnosis and/or treatmentcontaining the porphyrin compound represented by formula (I) or (II) aswell as a mixture thereof, or a pharmaceutically acceptable saltthereof.

[0019] More specific embodiment of the present invention, it is provideda photosensitizer for the photodynamic diagnosis and/or treatment oftumor of animals containing the porphyrin compound represented byformula (I) or (II), or a pharmaceutically acceptable salt thereof as anactive ingredient.

BRIEF DESCRIPTION OF DRAWINGS

[0020]FIG. 1 shows an infrared absorption spectrum of sodium salt of theporphyrin compound of the formula (I) (NOEt-P-Asp).

[0021]FIG. 2 shows the results of the accumulability to canceroustissues (cancer/organ concentration) of the porphyrin compound of theformula (I) (NOEt-P-Asp). In the graph, the curve No.1 represents theresult of cancer/brain, the curve No.2 represents the result ofcancer/liver, the curve No.3 represents the result of cancer/lung, thecurve No.4 represents cancer/muscle, the curve No.5 representscancer/kidney, and the curve No.6 represents cancer/plasma.

BEST MODE FOR CARRYING OUT THE INVENTION

[0022] The porphyrin compound of the present invention represented byformula (I) or (II) is a single component, is stable, and has a higherexcretion rate from normal tissues. Therefore, it is characterized thatthe porphyrin compounds of formula (I) or (II) has a reducedphototoxicity while retaining a good accumulability to cancerous tissuesand, furthermore, allows the use of titanium-sapphire laser (wavelengthof not less than 670 nm and not more than 600 nm) and a semiconductorlaser (wavelength of 670 nm).

[0023] Furthermore, when the porphyrin compound of formula (I) wasexamined by albumin test and dancyl methionine test, in which one of thepresent inventors has found a certain rule, it was confirmed that thecompound of formula (I) shows an excellent transferability to canceroustissues and a strong photosensitivity.

[0024] Albumin test is a test method for evaluating the affinity tocancerous tissues, in which a chlorin derivative is examined on thechange in ultraviolet (UV) absorption spectrum in a mixture form withalbumine, and dancyl methionine test is also a convenient test methodfor evaluating the strength of the photoreactivity by thin layerchromatography (TLC) or high performance liquid chromatography (HPLC)(see Japanese Patent Application Laid-open No. 5-97857).

[0025] The porphyrin compounds represented by formula (I) or (II) of thepresent invention can be prepared by the method as mentioned below.

[0026] That is, the compound can be prepared by a method comprising Step(a) in which a protoporphyrin dimethyl ester (hereinafter referred to as“PP-Me”), as starting compound, is converted into a chlorin derivativehaving an aldehyde group therein; Step (b) in which the aldehyde groupof the chlorin derivative thus obtained is converted to O-ethyliminogroup by condensation with a O-ethylhydroxylamine; and Step (c) in whichthe compound thus obtained is further introduced with aspartic acid viaan amide bond. It is not essential to conduct the reactions in the orderof (b) then (c), that is, compound of the present invention can beproduced in good yield in the case the compound is condensed withaspartic acid to form the amide bond as in Step (c), and then, thealdehyde group of the compound thus obtained is converted toO-ethylimino group by condensation with a O-ethylhydroxylamine as inStep (b).

[0027] Each of the steps is explained in more detail in the following.

[0028] Step (a) for conversion of the starting compound into a chlorinderivative can be conducted according to any of the conventionalmethods, such as methods disclosed in J. E. Falk: “Porphyrins andMetalloporphyrins” published by Elsevier in 1975; D. Dolphin: “ThePorphyrins” published by Academic Press in 1978 and so on.

[0029] That is, in step (a), PP-Me is subjected to a photochemicalreaction treatment to give 7-hydroxy-8-oxoethylidene-protoporphyrindimethylester (hereinafter referred to as “P-Me(I)”) and2-hydroxy-3-oxoethylidene-protoporphyrin dimethylester (hereinafterreferred to as “P-Me(II)”) in a mixture form. The later compound, i.e.P-Me(II), is a position isomer of P-Me(I) with respect to theside-chained substituent in the A and B rings of four tetrahydropyrrolerings. From the mixture thus obtained, each of P-Me(I) and P-Me(II) wasisolated and purified by means of silica gel column chromatography orrecrystallization using a suitable solvent. A mixture of P-Me(I) andP-Me(II) can be used for next step (b) without isolation.

[0030] Next, in step (b), the aldehyde group of P-Me(I), which isisolated and purified in step (a), is converted to O-ethylimino group bycondensation reaction with O-ethylhydroxylamine hydrochloride. Thisreaction can be conducted according to a conventional procedure asdisclosed in “Condensation reaction between hydroxylamine and analdehyde compound” in Ippan Yuki Kagaku Jikken Sho (Text for GeneralOrganic Chemical Experiments).

[0031] For example, the reaction may be conducted in suitable inertsolvent in the presence of condensation reagent such as inorganic ororganic base. Inorganic base may include alkali hydroxide or alkalimetal carbonate, and organic base may include pyridine or piperidine.The reaction can preferably be carried in pyridine or piperidine usingas a reaction solvent and as condensation reagent.

[0032] Accordingly, 7-hydroxy-8-ethoxyiminoethylidene-protoporphyrindimethyl ester (hereinafter referred to as “NOEt-P-Me(I)”) is convertedfrom P-Me(I), and P-Me(II), which is a position isomer of P-Me(I), isalso converted to 2-hydroxy-3-ethoxyiminoethyliden-protoporphyrindimethyl ester (hereinafter referred to as “NOEt-P-Me(II)”) in the samemanner.

[0033] Thus obtained NOEt-P-Me(I) is subjected to step (c). That is,NOEt-P-Me(I) is hydrolyzed with an alkali in a conventional manner andthen amidated with aspartic acid methyl ester to obtain aspartic acidsubstituted porphyrin.

[0034] This reaction may be conducted by a conventional procedure asdisclosed in Izumiya et al.,: “Peptide gosei no kiso to jikken(Basis andExperiments of Peptide Synthesis)”, published by Maruzen in 1985, andespecially a procedure as disclosed in Japanese Patent ApplicationLaid-open Nos. 64-61481, 2-138280, 4-59779, 5-97857 or 9-124652, orJapanese Patent Publication No. 7-25763 are preferred.

[0035] By this reaction, aspartic acid residue is introduced in the sidechain of the porphyrin compound, and the reaction may occur betweencarboxyl group at the side chain of porphyrin compound and amino groupof aspartic acid. Therefore, it should be considered in the reaction toconvert the carboxyl group at the side chain of porphyrin compoundand/or amino group of aspartic acid to reactive substituent byconventional manner, or to protect functioning group not preferable toparticipate in both groups.

[0036] The reaction may be accelerated in suitable solvent usingreaction accelerator such as dehydration agent and deoxidation agentwhich examples are dicyclohexylcarbodiimide (DCC) and water solublecarbodiimide (WSC).

[0037] According to above reaction, NOEt-P-Me(I) for example, isamidated with aspartic acid dimethylester after alkali hydrolysis, andthen derived to 7-hydroxy-8-ethoxyiminoethiliden-protoporphyrin[hereinafter referred to as “NOEt-P-Asp(OMe)(I)].

[0038] In the same manner, NOEt-P-Me(II) is converted to2-hydroxy-3-ethoxyiminoethiliden-protoporphyrin [hereinafter referred toas “NOEt-P-Asp(OMe)(II)].

[0039] NOEt-P-Asp(OMe)(I) or NOEt-P-Asp(OMe)(II) thus obtained ishydrolyzed by, for example, sodium hydroxide after dissolved andsuspended in, for example, ethanol, thus sodium salts of the porphyrincompounds of the present invention represented by formula (I) or formula(II) is obtained.

[0040] Free carboxylic acid of the porphyrin compound is derived fromtreating these sodium salts with weak acid.

[0041] Accordingly, compounds stated below are provided as porphyrincompounds of the present invention.

[0042] (1)13,17-bis[(1,2-dicarboxylethyl)carbamoylethyl]-3-ethenyl-7-hydroxy-8-ethoxyiminoethylidene-2,7,12,18-tetramethyl-porphyrin[hereinafter referred to as “NOEt-P-Asp(I)”],

[0043] (2)13,17-bis[(1,2-dicarboxyethyl)carbamoylethyl]-8-ethenyl-2-hydroxy-3-etoxyiminoethylidene-2,17,12,18-tetramethyl-porphyrin(hereinafter referred to as “NOEt-P-Asp(II)”).

[0044] The porphyrin compound provided by the present invention is usedfor a photodynamic diagnosis and/or treatment of animals. Formulation ofthe compound is done according to the common method to the ones skilledin the art. When the porphyrin compound of the present invention is freeacid, the object agent is formulated by dissolving it in suitablebuffer, whereas in the case the porphyrin compound of the presentinvention is sodium salt, the object agent is formulated by dissolvingit in physiological saline. Examples of suitable additives to be usedare pharmaceutically acceptable adjuvant such as organic solvent, pHadjuster such as acid, base and buffer, stabilizer such as ascorbicacid, excipient such as glucose and isotonic agent such as sodiumchloride.

[0045] The porphyrin compound of the present invention possessesfeatures of photosensitizer for PDT such as long phosphorescence life,remarkable accumulability to specific internal organ, especially tocancer locus, good cell killing effect when exposed to light asdetermined by a dancyl methionine test, excellent absorption wavelength,water solubility and purity.

[0046] The good water solubility of this compound enables a preparationof a high concentration solution such as 50 mg/ml. Furthermore, thecompound exhibits a high stability in vivo, as well as in vitro. Whenused for photodynamic diagnosis and/or treatment of animals asphotosensitizer for PDT in general, it is desirable to administer thecompound to a subject in a dose of 1-10 mg/kg body weight.

[0047] As discussed above, the porphyrin compound of the presentinvention is structurally characterized in that it has an amino acidresidue, especially aspartic acid residue, and further ethoxyliminogroup, and as result, it exhibits various physiological andpharmacological properties.

[0048] As one of the properties, the compound selectively accumulates intumor cells and is excreted therefrom at a slow rate. On the other hand,excretion from normal organs and cells is rapid and therefore it doesnot damage such organs and cells, and does not cause phototoxicity.

[0049] Furthermore, according to the present invention, the conversionof a porphyrin into a chlorin derivative allows the absorptionwavelength to shift to infrared region and, as a result, it becomespossible to attain therapeutic efficacy for cancers in deep site.Accordingly, the porphyrin derivative of the present invention is highlyuseful as a photosensitizer for PDT for cancers and malignant tumors inanimals.

EXAMPLES

[0050] The present invention will be described in more detail byreferring to the following examples.

Example 1 Synthesis of Mixture of7-hydroxy-8-oxoethylidene-protoporphyrin dimethylester [P-Me(I)] and2-hydroxy-3-oxoethylidene-protoporphyrin dimethyl ester [P-Me(II)],Position Isomer of P-Me(I)

[0051] The title compounds were synthesized by the method of P. K.Dinello et al., (“The porphyrins”, academic press, Vol. 1, 303(1978)).100 g of protoporphyrin dimethyl ester (pp-Me) was dissolved in 10 L ofchloroform. The resultant reaction mixture was allowed to react for oneweek under irradiation with light, thereby obtaining a mixture ofchlorin derivatives of the porphyrin. After completion of the reaction,the reaction solution was concentrated under reduced pressure to givethe residue (100 g) containing the title compounds in the mixture form.

Example 2 Isolation of P-Me(I) and P-Me(II) from Resultant Mixture

[0052] The resultant mixture obtained in Example 1 was dissolved in amixture solution of dichloromethane-hexane, and the mixture solution wassubjected to silica gel chromatography to eliminate insoluble matter,then the filtrate was concentrated. The resultant residue was treatedwith ethyl acetate, and obtained solid was recrystallized withpyridine-dichloromethane to give7-hydroxy-8-oxoethylidene-protoporphyrin dimethylester [P-Me(I)].Further, 2-hydroxy-3-oxoethylidene-protoporphyrin dimethylester[P-Me(II)], position isomer of P-Me(I), was obtained from therecrystallized filtrate.

Example 3 O-ethylimination and Hydrolysis of P-Me(I) and P-Me(II)

[0053] Each of P-Me(I) and P-Me(II) obtained in Example 2 was separatelyweighed out (10 g each) and dissolved in pyridine (190 ml,)respectively. To the resultant solution was added 3 g ofO-ethylhydroxylamine hydrochloride and allowed to react at 50° C. for1.5 hours under stirring. After the reaction was completed, the reactionsolution was poured into water to precipitate a crystalline substance.The crystalline substance was collected by filtration. In this manner,O-ethylimino P-Me(I) [NOEt-P-Me(I)] and O-ethylimino P-Me(II)[NOEt-P-Me(II)] were obtained (yield: quantitative).

[0054] Each of NOEt-P-Me(I) and NOEt-P-Me(II) obtained in the aboveprocedure was dissolved in pyridine (200 ml) separately. The resultantsolution was hydrolyzed with 1N sodium hydroxide solution in aconventional manner, and the reaction solution was neutralized, therebygiving a precipitate. The precipitate thus obtained was collected byfiltration, washed and dried, and further purified with ethylacetate-hexane mixture solution to give NOEt-P(I) and NOEt-P(II) (yield:quantitative).

Example 4 Conversion of NOEt-P(I) and NOEt-P(II) into Aspartic AcidDerivatives Thereof

[0055] (a) Each of NOEt-P(I) and NOEt-P(II) obtained in Example 3 wasseparately weighed out (2 g each), dissolved in tetrahydrofuran, andconverted into a dicyclohexylamine (DCHA) salt (2.0 g each) with DCHA ina conventional manner, respectively.

[0056] (b) Each of the resultant DCHA salts was dissolved indimethylformamide. To the resultant solution was added aspartic aciddimethyl ester (AspOMe₂) hydrochloride and further added water solublecarbodimide (WSC). Each of the resultant solutions was allowed to react.After confirming the completion of the reaction by TLC, water was addedto each reaction solution to thereby cause precipitation. Each resultantprecipitate was washed with water, dried and dissolved in acetone/ethylacetate mixture solution. The resultant mixture solution was, thenpurified by silica gel chromatography, and recrystallized to obtainNOEt-P-Asp(OMe)(I) and NOEt-P-Asp(OMe)(II) as dark greenish browncrystals.

Example 5 Production of NOEt-P-Asp(I) and NOEt-P-Asp(II)

[0057] Each of NOEt-P-Asp(OMe)(I) and NOEt-P-Asp(OMe)(II) obtained inExample 4 was separately weighed out (1 g each) and hydrolyzed in aconventional manner by dissolving them in ethanol and then 1N sodiumhydroxide. After the reaction was completed (The reaction end point wasconfirmed by TLC.), ethanol was added to each reaction solution tothereby cause precipitation. Each resultant precipitate was collected byfiltration and dissolved in water. To each resultant solution was addedadditional ethanol to thereby cause precipitation for furtherpurification. In this manner, a sodium salt of NOEt-P-Asp(I) wasobtained from NOEt-P-Asp(OMe)(I), and a sodium salt of NOEt-P-Asp(II)was obtained from NOEt-P-Asp(OMe)(II). MS:955 (M⁺)

[0058] The infrared absorption spectrum of the sodium salt ofNOEt-P-Asp(I) is shown in FIG. 1.

Example 6 Evaluation on Tissue Accumulability

[0059] 3H/He mice (5 per group) implanted with tumor tissues of coloncancer Colon 26 for 14 to 21 days, were given an intravenous injectionof sodium salt of each of NOEt-P-Asp(I) (10 mg/kg for each mouse) whichhad been diluted with a distilled water for injection. The blood sampleswere collected and the organs bearing the tumor tissue were extirpatedafter the injection, irradiated with N₂-pulsed laser (N₂, wavelength:337 nm, 2 ns, 400-1,000 nm), and the excited fluorescent spectrum wasmeasured. The wavelength in the range of 600 to 900 nm was examinedbased on the peak wavelength of NADH at 470 nm (determination of thedistribution of the test compound in the organ by the surfacefluorescence method using N₂-pulsed laser spectrophotometry). That is,the distributed concentration of sodium salt of NOEt-P-Asp(I) incancer/organ (or plasma) ratio was determined by calculating the peakwavelength at 670 nm when the peak wavelength at 470 nm was consideredas the basic value, 1. The result obtained after administration 1 to 24hours is shown in FIG. 2. The sodium salt of NOEt-P-Asp(I) was found tohave much higher accumulability to cancerous tissues.

Example 7 Evaluation on Photosensitizing Oxidation Reaction Using DansylMethionine

[0060] To 1 ml of chloroform, 10 μM of Dansyl methionine, a substrate,was added, and then 0.1 μM of the photosensitizer of the presentinvention [sodium salt of NOEt-P-Asp(I)] was further added. Laserirradiation was conducted using Cold Spot PICL-SX (Nippon P. I. Co.,Ltd.) which is halogen lamp, has 150 W wave length and 80,000 Lux, understirring. The reaction solution was spotted at every minute on TLC plate(Kieselgel 60F254), and developed with chloroform methanol (3:2), thenDansyl methionine and its oxide (Dansylmethionine sulfoxide) wereconfirmed using UV lamp (254 nm). The time that Dansylmethioninedisappeared completely on TLC plate was stated as the end of thereaction time, and photo oxidation reaction of photosensitizer wascompared.

[0061] Photofrin II (Trade Mark) was used as a control ofphotosensitizer of present invention.

[0062] The result was shown in Table 1 below. The values in the tableshow the reaction end time in minutes, thus it shows that the smallervalues (minutes) is, stronger the photosensitizing reaction.

[0063] As clearly shown in the table, photosensitizing agent of thepresent invention shows stronger photosensitivity reaction compared withPhotofrin II (Trade Mark). TABLE 1 Name of the Compound Degree of PhotoReaction Photofrin II 10< NOEt—P-Asp(I) Na salt 4 NOEt—P-Asp(II) Na salt4

[0064] Industrial Applicability

[0065] The porphyrin compound of the present invention has a highaccumulability to cancerous cells, reactivity to external energy and acancerous cell destroying effect. Furthermore, it exhibits no toxicityagainst normal cells. Accordingly, it is extremely useful as adiagnostic and therapeutic agent for cancer for animals.

1. A porphyrin compound represented by the following formula (I):wherein Asp represents a residue aspartic acid, or a pharmaceuticallyacceptable salt thereof, used for photodynamic diagnosis and/ortreatment of animals.
 2. The photodynamic diagnostic and/or therapeuticagent on cancers for animals comprising the porphyrin compound offormula (I) or pharmaceutically acceptable salt thereof according toclaim 1 as an active ingredient.
 3. The photodynamic diagnostic and/ortherapeutic agent claimed in claim 2, which is used for photodynamicdiagnosis and/or treatment on cancers for animals.
 4. A porphyrincompound represented by the following formula (II): wherein Asprepresents a residue of aspartic acid, or a pharmaceutically acceptablesalt thereof, used for photodynamic diagnosis and/or treatment ofanimals.
 5. The photodynamic diagnostic and/or therapeutic agent oncancers for animals comprising the porphyrin compound of formula (II) orpharmaceutically acceptable salt thereof according to claim 4 as anactive ingredient.
 6. The photodynamic diagnostic and/or therapeuticagent claimed in claim 4, which is used for photodynamic diagnosisand/or treatment on cancers for animals.
 7. The photodynamic diagnosticand/or therapeutic agent for animals comprising the mixture substance ofthe porphyrin compound of formula (I) claimed in claim 1 and theporphyrin compound of formula (II) claimed in claim 2 orpharmaceutically acceptable salt thereof as an active ingredient.
 8. Thephotodynamic diagnostic and/or therapeutic agent claimed in claim 7,which is used for photodynamic diagnosis and/or treatment on cancers foranimals.